4 (1-2)  December 2013: 87-92 Devetaković, J.,et al.  Variability of morphometric caracteristic of… BI OLOGICA NYSSANA

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Original Article

Variability of morphometric caracteristics of one-year seedlings of different half-sib European White Elm (Ulmus effusa Wild.) from the

Jovana Devetaković, Mirjana Šijačić-Nikolić, Vladan Ivetić

Faculty of Forestry, , Kneza Višeslava 1, 11000 Belgrade,

* E-mail: [email protected]

Abstract: Devetaković, J., Šiljačić-Nikolić, M., Ivetić, V.: Variability of morphometric caracteristics of one-year seedlings of different half-sib European White Elm (Ulmus effusa Wild.) from the Great War Island. Biologica Nyssana, 4 (1-2), December 2013: 87-92. European White Elm is recognized as a rare and endangered species in the forest fund of the Republic of Serbia. During the past century massive drying of elms occurred and the indications of their extinction appeared, which consequently led to a reduction in genetic diversity and the danger of genetic drift. In the area of the Great War Island near Belgrade we found 56 trees of European White Elm that are spatially divided into three subpopulations. In order to assess the genetic potential of European White Elm in the Great War Island and to define adequate conservation measures variability of 14 selected test trees progeny was rated. Results shows that the satisfactory variability within the popoulation exists, which is a good basis for the conservation of genepool available. Key words: conservation, European White Elm, genepool, seedlings, Ulmus effusa, variability.

Introduction main cause of the often drying of these species. However, disappearance of wetlands due to drainage European White Elm (Ulmus effusa Wild., for agricultural production or poplar cultivation is syn. U. laevis Pall.) is one of the three more often considered to be the cause of European autochthonous species of familiy Ulmaceae Mirb. in White Elm drying (Collin 2003). Serbia. Unlike Field Elm (Ulmus minor Miller) and In the forest fund of the Republic of Serbia, Mountain Elm (Ulmus montana With.), which European White Elm is recognized as a rare and occupy slightly different habitats, European White endangered species (B a n k o v i ć et al. 2009). From Elm (Ulmus effusa Wild.) is a species limited to the the perspective of conservation and directed lowlands. Looking outside of Serbia, European utilization of forest genetic resources, European white elm grows in Central and Eastern Europe White Elm can be considered as priority species in (J o v a n o v i ć 2007). sence of the available genepool, and the fact that During the past century massive drying of conservation areas for in situ conservation of this elms occurred and the indications of their extinction species in Serbia do not exist, and the methods of ex appeared, both in natural populations and in urban situ conservation were not performed (Š i j a č i ć - areas, which consenquently lead to the reduction of N i k o l i ć , M ilovanović 2010). genetic diversity and the danger of genetic drift. The variability and growth of European White Dutch elm disease (DED) is considered to be the Elm seedlings were studied by many authors.

87 BIOLOGICA NYSSANA 4 (1-2)  December 2013: 87-92 Devetaković, J.,et al.  Variability of morphometric caracteristic of…

B l a c k - S a m u e l s s o n et al. (2003) reported existence of a strong additive genetic variation between and within populations in growth of one- year old seedlings as response to drought stress. W i t h e l e y et al. (2003) also reported iner- and intrapopulation variability of one-year old seedlings. Examining of the four-year old plants of Fraxinus angustifolia and Ulmus laevis, C i c e k et al. (2011), reports influence of provenance and growth density on the development of ash seedlings, but lack of significant influence on European White Elm seedlings.

Material and methods

A total of 56 trees of Europen White Elm were found, marked and georeferenced on Great War Island, near Belgrade, Serbia (Fig. 1). The trees are spatially divided into three subpopulations: Fig. 1. Spatial distribution of subpopulations European White Elm (Ulmus effusa Willd.) at Great War Island

Table 1. Coordinates and basic characteristics of 14 mother trees of European White Elm (Ulmus effusa Willd.) at Great War Island Mark of Coordinates Average Height Subpopulation tree on diameter Age E N (m) the field (cm) 13 7456215 4965040 23,1 48.7 26 I 14 7456215 4965038 7,7 15.65 13 32 7456352 4964955 20,2 38.3 23 18 7456050 4965266 16,7 39.5 27 19 7456054 4965270 18,4 36.5 16 21 7456046 4965285 9,5 31.5 16 II 33 7456138 4965226 13,7 29.35 20 34 7456170 4965254 21 31.3 20 35 7456150 4965257 20,9 30.4 20 36 7456136 4965262 13,9 37.8 23 30 7455319 4965639 13,5 23.5 15 III 31 7455137 4965585 12 23.5 17

The first subpopulation (I) consists of 24 trees development of floodplain forests of soft deciduous located in the inshore area, the recent alluvial trees. European White Elm (Ulmus effusa Willd.) is deposits with characteristics of β-gley, White seen here in the second floor of the trees, beside Willow forest type (Salicetum albae Issl.26) on the Black poplar (Populus nigra L.) and White poplar β -gley. (Populus alba L) edificators (Š i j a č i ć - N i k o l i ć , The second subpopulation (II) consists of 22 Milovanović 2012). marked trees inhabiting the area with a very dense In the spring of 2011, during May, the seeds canopy and impervious. It extends in the internal were collected from all trees where yield was part of the island, close to the canal Galijaš. recorded (Tab. 1). The third subpopulation (III) consists of 10 Seeds were collected in May 2011 and after a trees individually dispersed throughout the forest short drying during the same month were sown in edge or in a field. The second and the third seed beds, seeding density 1200 seeds/m2. subpopulation belong to the forest of White and Seed beds were covered with a thin layer of Black poplar (Populetum albo-nigra Slav.52) the the substrate made up of sand and soil in the ratio of mosaic of alluvial soil, which is the final stage of 1:1 (Fig. 2 and Fig. 3). Irrigation was initially done

88 BIOLOGICA NYSSANA 4 (1-2)  December 2013: 87-92 Devetaković, J.,et al.  Variability of morphometric caracteristic of…

Basic statistical analysis of the data collected included the calculation of mean values, standard deviations (SD) and width variation. Clustering of the obtained mean values and the homogenity of groups were tested by Tukey HSD test. The origin of variability was examined by analysis of variance (one-way ANOVA). The interdependence of the morphological parameters was tested by calculating the Pearson correlation coefficient. Closeness (distance) of mother trees based on the above parameters was tested by cluster analysis.

Fig. 2. Seed beds with seed partialy covered daily until seedlings appeared, and then less frequently (every 3-4 days) (Figure 4). Each of the half-sib lines samples was taken from 50 one-year plants with bare-roots, their height (H) and root collar diameter (D) were measured. Roller’s sturdiness coefficient, as the ratio of the height (cm) and diameter (mm), were calculated (R o l l e r 1977, I v e t i ć 2013).

Fig. 4. Appearance of seedlings after germination

Fig. 3. View on the seed beds sown and covered Fig. 5. The layout of rows of seedlings

89 BIOLOGICA NYSSANA 4 (1-2)  December 2013: 87-92 Devetaković, J.,et al.  Variability of morphometric caracteristic of…

Table 2. Mean values of height (H), diameter (D) and Roller’s sturdiness coefficient (SQ) (standard deviation (SD in parentheses), the limits for H, D, SQ and HSD test of 14 tested maternal line (ML)

N=700 H D SQ MEAN MEAN MEAN ML X -X X -X X -X VALUES (Sd) mиn max VALUES (Sd) mиn max VALUES (Sd) mиn max 13 20.55 (5.92)abc 12.3-40.2 3.25 (1.07)abc 1.47-7.05 6.52 (1.35)a 3.73-10.1 14 19.34 (5.64)ab 10.4-36.5 3.13 (1.17)abc 1.43-6.49 6.45 (1.29)a 3.98-9.5 18 23.34 (6.44)abc 12.2-41.2 3.46 (1.31)abc 1.67-6.57 7.18 (1.81)abc 3.81-10.6 19 23.83 (8.13)bc 9.3-46.8 3.42 (1.59)abc 1.30-7.69 7.52 (1.97)abc 4.24-15.8 21 18.87 (7.55)a 7.9-45.0 2.80 (1.35)a 0.88-7.33 7.12 (1.48)ab 4.40-10.4 29 21.93 (7.29)abc 11.0-43.4 3.52 (1.35)abc 1.55-6.97 6.51 (1.44)a 3.25-10.3 30 29.89 (9.33)d 13.4-55.7 3.91 (1.66)c 1.79-8.86 8.29 (2.80)c 2.84-21.8 31 21.67 (8.87)abc 8.9-47.1 2.91(1.35)ab 1.17-6.97 7.71 (1.53)bc 4.44-11.5 32 20.33 (5.88)abc 10.6-35.8 2.89 (0.96)ab 1.48-5.97 7.25 (1.50)abc 4.03-11.1 33 24.92 (7.94)c 7.0-43.7 3.73 (1.32)bc 1.31-7.84 6.87 (1.53)ab 4.52-10.9 34 24.73 (7.54)c 10.5-43.5 3.45 (1.34)abc 1.11-7.96 7.53 (1.64)abc 3.68-11.1 35 23.41 (6.27)abc 7.8-38.7 3.31 (1.04)abc 1.73-6.00 7.36 (1.98)abc 3.50-12.1 36 21.84 (5.94)abc 11.1-35.0 3.29 (0.98)abc 1.36-5.24 6.88 (1.45)ab 4.18-9.9 37 20.48 (7.56)abc 6.5-35.4 2.80 (1.35)a 0.97-6.96 7.86 (1.91)bc 3.46-11.6 ALL 22.5 (7.70) 6.5-55.7 3.28 (1.32) 0.88-8.86 7.22 (1.79) 2.84-21.8 LINES

Results and discussion Table 3. Analysis of variance (one-way ANOVA) height (H) and diameter (D) of 14 half-sib lines of The highest average H (29.89 cm) and D European White Elm (3.91 mm) was observed in half-sib seedlings line 30 of the parent tree (Table 2). The maximum BETWEEN MATERNAL LINES values of 55.7 cm H and D of 8.86 mm were СВОЈСТ MEAN P- F-RATIO recorded in the same line of seedlings at the same ВО SQUARE VALUE time. H 40721.00 7.74 0.00 Mean values in the same column followed by D 1.67 3.48 0.00 different letters are statistically different for p <0.05. According to the observed parameters seedlings from 30 lines segregate into a separate Table 4. Correlations between the morphological group. They also show the highest variability of parameters: the diameter (D), the height (H) and observed parameters. Thus, for H Sd = 9.33 with Roller’s sturdiness coefficient (SQ) of 14 half-sib variational width of 42.30 cm for D Sd = 1.66 with lines of European White Elm the variation width of up to 7.07 mm, and for SQ Sd = 2.8 the variation width of 18.96. N=700 (Casewise deletion of missing data) Minimum value of 6.5 cm H and D of 0.97 D H SQ mm were measured in seedlings of maternal line 37, D 1.00000 while the minimum mean values of both measured H 0.79095* 1.00000 parameters were recorded in the line 21. Analysis of variance indicated that the SQ -0.48323* 0.10136* 1.00000* variability of physiological parameters is the consequence of a mother tree, not a coincidence, Figure 6 shows a constant growth of SQ with and that the variability between seedlings of increasing of H. On the other hand, the value of SQ maternal lines (p = 0.00) is greater than the decreases with growth of D, to certain, variability of plants within a maternal line (Table 3). approximately mean values. After that, with the D and H showed a significant and strong increase of D, SQ begins to grow. positive correlation. SQ shows a relatively strong, negative correlation with D, and a weak, but significant correlation with H.

90 BIOLOGICA NYSSANA 4 (1-2)  December 2013: 87-92 Devetaković, J.,et al.  Variability of morphometric caracteristic of…

For the purpose of ex situ 3D Surface: H vs. D vs. SQ conservation, the level of variation (Casewise deletion of missing data) within the population should be Z = Distance Weighted Least Squares compared with the close spatial populations of European White Elm of coastal areas of the Danube and Sava rivers, during their work, W h i t e l e y et al. (2003) found that studied populations differ in the level of genetic variation within the population. Also, exploring the genetic variability of the border populations of European White Elm, V a k k a r i et al. (2009) suggest that differences between populations follow the structure of isolation 40 caused by distance (isolation by 30 distance). 20 10 0 Conclusion

The vulnerability of European Fig. 6. Area of a combination of height (H) and diameter (D) as a White Elm (Ulmus effusa Wild.) predictor, as opposed to Roller’s sturdiness coefficient (SQ) as the population on the Great War Island, dependent variable is sufficiently explained by the record that only 56 trees of this Cluster analysis (Fig. 7) shows clustering of species exist, and their age structure indicates the examined maternal lines into three groups. The first existence of different generations of trees in the group consists of the maternal lines 18, 19, 33, 34 population. and 35. The second group consists of lines 13, 14, According to the research of variability of 21, 29, 31, 32, 36 and 37. The third group consists morphometric caracteristics of one-year old of 30 lines, which are groups at the greatest distance seedlings, it can be concluded that the satisfactory with other maternal lines. variability within the popoulation exists, which is a The density of growth, as a result of planting good basis for the conservation of genepool density has a strong effect on the growth of available. Special attention should be paid to the seedlings. Applied seeding density of 1,200 seeds half-sib lines whose characteristics (height, diameter per m2 significantly exceeds the growth of 75 plants and Roller’s sturdiness coefficient) provide the best per m2, where C i c e k et al. (2011) found the quality seedlings, which in this case is the line half- highest values of H and D of one-year old seedlings sib seedlings of mother tree 30. But from the aspect of European White Elm with bare-root. However, it of conservation and directed utilization the specific did not significantly affect the SQ, which is genotypes which manifested the minimum satisfactory and which is close to the results got by dimensions such as tree seedlings of lines 21 and 37 the listed authors. SQ change with increasing D should not be ignored. should be explored, especially in the second year of For effective protection of genetic resources growth of seedlings. of European White Elm on the Great War Island, in Variability within the population of situ and ex situ methods should be combinated for a parameters observed is significant, as shown by the long time. In situ conservation is already partially analysis of variance, and does not depend on the realized, given that it is a protected nature reserve. spatial distribution of mother trees. To apply the ex situ methods it is necessary to Considering the number of trees in the continue monitoring of half-sib lines and establish reference population as an appropriate method may progeny tests, in order to select appropriate trees for be recommended the in situ conservation of genetic the production of plants and to expand the species resources. In endangered populations of European population and conservation of her genetic White Elm, with a significantly reduced number of variability in this area. trees, may be recommended the ex situ conservation (Aleksic, Orlovic 2004).

91 BIOLOGICA NYSSANA 4 (1-2)  December 2013: 87-92 Devetaković, J.,et al.  Variability of morphometric caracteristic of…

Tree Diagram for 14 Cases Single Linkage Euclidean distances

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0 10 20 30 40 50

Linkage Distance

Fig. 7. Dendrogram of the cluster analysis was done on the basis of height and root collar diameter of one- half-sib seedlings of European White Elm

References Jovanović, B. 2007: Dendrologija. Šumarski fakultet Univerziteta u Beogradu. Str. 1-536ultet, Aleksić, J., Orlović, S. 2004: Ex situ conservation of Beograd: 1-77. genetic resources of field elm (Ulmus minor Ivetić, V. 2013: Praktikum iz Semenarstva Mill.) and european white elm (Ulmus laevis rasadničarstva i pošumljavanja. Univerzitet u Pall.). – Genetika, Vol. 36, No. 3, 221-227. Beogradu, Šumarski fakultet, 213 p.. Banković, S., Medarević, M., Pantić, D., Petrović, Šijačić-Nikolić, M., Milovanović, J. 2010: N., Obradović, S. 2009: Šumski fond Republike Konzervacija i usmereno korišćenje šumskih Srbije-stanje i problemi, Glasnik Šumarskog genetičkih resursa, Šumarski fakultet fakulteta, Beograd, br. 100, (7-30). Univerziteta u Beogradu, Beograd, 1-200. Collin, E. 2003. EUFORGEN Technical Guidelines Šijačić-Nikolić, M., Milovanović, J. 2012: for genetic conservation and use for European Conservation and sustainable use of foerst white elm (Ulmus laevis). International Plant genetic resources throught an example of Genetic Resources,Institute, Rome, Italy. 6 p. watland ecosystems, International Conference: Black-Samuelsson S., Whiteley R. E., Junzhan G. Role of research in sustainable development of 2003: Growth and Leaf Morphology Response agriculture and rural areas, May 23-26, to Drought Stress in the Riparian Broadleaved Podgorica, Montenegro, Book of Abstracts, 128. Tree, Ulmus laevis (Pall.). Silvae Genetica 52, Whiteley, R.E., Black-Samuelsson, S., Jansson, G. 5-6. 2003: Within and between population variation Cicek E., Cicek N., Tilki F. 2011: Four-year Field in adaptive traits in Ulmus laevis Pall., the Performance of Fraxinus angustifolia Vahl. and European white elm. Forest Genetics 10(4):313- Ulmus laevis Pall. Seedlings Grown at Different 323. Nursery Seedbed Densities. Research Journal of Forestry, 5: 89-98.

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